摘要
目的:探讨SOX4与食管癌顺铂耐药的关系及其耐药机制。方法:采用递增药物浓度、间歇冲击作用的方法构建顺铂食管癌耐药细胞株KYSE30/DDP;流式细胞仪检测KYSE30及KYSE30/DDP细胞凋亡情况;Western blot检测SOX4及凋亡相关指标在KYSE30及KYSE30/DDP细胞中的变化;CCK8检测干扰SOX4对KYSE30和KYSE30/DDP细胞顺铂敏感度的影响;流式细胞仪及Western blot检测干扰SOX4对KYSE30/DDP细胞凋亡及相关指标的影响。结果:成功构建了生长状态良好的KYSE30/DDP细胞株,其耐药指数为4.1;KYSE30/DDP细胞凋亡较其亲本细胞KYSE30明显降低(t=-8.414,P<0.05);Western blot结果显示,SOX4在顺铂耐药细胞KYSE30/DDP中的表达高于其亲本细胞KYSE30,KYSE30/DDP细胞中凋亡相关指标Cleaved caspase-3及Cleaved PARP表达低于KYSE30;在KYSE30/DDP细胞中干扰SOX4,其对顺铂的敏感性及凋亡细胞增加(F=625.423,P<0.05),凋亡相关指标Cleaved caspase-3及Cleaved PARP表达增加。结论:SOX4在食管癌顺铂耐药细胞KYSE30/DDP中高表达,并可通过抑制凋亡介导食管癌顺铂耐药。
Objective：To explore the relationship between cisplatin resistance and the expression of SOX4, and related drug resistance mechanisms in esophagus cancer. Methods：Thecisplatin resistance cell line KYSE30/DDP was established in culture by a program of treating KYSE30 parent cells in culture with increasing concentration of cisplatin. The apoptosis of KYSE30 and KYSE30/DDP cells were determined by flow cytometry. Western blot detected the expression of SOX4 and apoptosis related indicators in KYSE30 and KYSE30/DDP cells. The influence of drug sensitively by knockdown the expression of SOX4 in KYES30 and KYSE30/DDP cell was measured by CCK8 assay.The change of apoptosis and related indicators in KYSE30/DDP cell after knockdown the expression of SOX4 was evaluated by western blot and flow cytometry.Results CCK8 assay showedthe cisplatin resistance cell line KYSE30/DDP was successfully built, and the resistance index was 4.1. The apoptosis of KYSE30/DDP cell was significantly decreased compared with KYSE30 parent cell（P〈0.05）. Western blot indicated the expression of SOX4 are higher in KYSE30/DDP cell compared to its parent cell. And the expression of apoptosis related indicators lower in KYSE30/DDP cell. Knock down the expression of SOX4 in KYSE30/DDP cellfacilitated thecisplatin sensitivity andpromoted cell apoptosis（P〈0.05）,with the increase expression of apoptosis related indicators . Conclusion SOX4 was high-expression in cisplatin resistance cell KYSE30/DDP, and mediated cisplatin resistance by inhibiting apoptosis.
出处
《肿瘤预防与治疗》
2017年第3期158-163,共6页
Journal of Cancer Control And Treatment
基金
泸州市政府-西南医科大学联合项目(编号:2013LZLY-J40)
关键词
食管癌
顺铂
SOX4
耐药
Esophagus Cancer
SOX4
Apoptosis
Cisplatin Resistance
